the need for rationale explanations of homeopathyppt – 3.5. 12 histamine dilutions modulate...

1

THE NEED FOR RATIONALE EXPLANATIONS OF HOMEOPATHY

• As reported by Kleijnen et al. in their seminal review on clinical trials in homeopathy*:

•

* Kleijnen, J., Knipschild, P. & Ter Riet, G. 1991. Clinical trials of homoeopathy. Brit. Med. J. 302: 316-323.

• It is only through patient, unrestricted, and methodical research conducted on several planes - clinical, laboratory, epidemiological, and physicochemical - that we shall be able to shed light on the many issues which so far remain unsolved.

- 3.2. The logic of the simile -*

2

LA TRIADE OMEOPATICA E LA RICERCA DI BASE

PPT PPT –– 2.1 2.1

ALTE POTENZEALTE POTENZEENERGIAENERGIA

BASSE POTENZEBASSE POTENZEMATERIAMATERIA

Comefunziona?

Comefunziona?

SIMILITUDINESIMILITUDINEDILUIZIONE/DILUIZIONE/

POTENZAPOTENZA

INDIVIDUALITAINDIVIDUALITA ’’

GLOBALITAGLOBALITA ’’

ORMESIORMESI

VALO

RE

INIZ

IALE

VALO

RE

INIZ

IALE

COMPLESSITACOMPLESSITA’’INFORMAZIONEINFORMAZIONE

BIOFISIC

A

BIOFISIC

A--

BIOELETTR

OM

AGN

ETISMO

BIOELETTR

OM

AGN

ETISMO

Comefunziona?

Comefunziona?

SIMILITUDINESIMILITUDINEDILUIZIONE/DILUIZIONE/

POTENZAPOTENZA

INDIVIDUALITAINDIVIDUALITA ’’

GLOBALITAGLOBALITA ’’

ORMESIORMESI

VALO

RE

INIZ

IALE

VALO

RE

INIZ

IALE

COMPLESSITACOMPLESSITA’’INFORMAZIONEINFORMAZIONE

BIOFISIC

A

BIOFISIC

A--

BIOELETTR

OM

AGN

ETISMO

BIOELETTR

OM

AGN

ETISMO

LABORATORIOe ANIMALI



CCC

AAA

BBBEEE

DDD

OMEODINAMICAOMEOPATOLOGIA

OMEOTERAPIACCC

AAA

BBBEEE

DDD


OMEOTERAPIACCC

AAA

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OMEOTERAPIA

FISICADELL’ACQUA,BIOELETTRO-MAGNETISMO,

ECC.


ECC.


ECC.


ECC.

3

Effects of homeopathic treatment on animal models of cancerogenesis (part 1)

*The formula of Canova is composed of 19x Thuya occidentalis, 18x Bryonia alba, 11x Aconitum napellus, 19x Arsenicum album and 18x Lachesis muta(Viperidae) venom, all extracted and diluted in 70% alcohol, in equal parts.

(Sato et al. 2005) Homeopathy 94 (1):26-32.

Delay in the development, increased infiltration by lymphoid cells with active treatment. Increased number of leukocytes and lymphocytes.

Canova, a homeopathic complex medicine*

Sarcoma 180Mice

(Biswas et al. 2005) J Altern.Complement Med 11 (5):839-854.

Carcinosin 200 and Chelidonium 200 when administered alone show considerable ameliorative effect on cytogenetical endpoints and toxicity biomarkers

Carcinosin 200, fed alone and in combination with Chelidonium 200

p-DAB-induced hepatocarcinogenesis

Mice

(Datta, Biswas, and Khuda-Bukhsh 2004) Evid.Based.Complement Alternat.Med 1 (3):291-300.

Less chromosome aberrations in the drug-fed series. The amelioration by Merc Sol-200 appeared to be slightly more pronounced.

MercuriusSolubilis(Merc Sol-30 and Merc Sol-200)

Genotoxic Effects Produced by Mercuric Chloride

Mice

(Biswas and Khuda-Bukhsh 2004) Indian J.Exp.Biol. 42 (7):698-714.

Both Ch-30 and Ch-200 also modulated favourably some toxicity marker enzymes

Chelidonium-30 (Ch-30) and Chelidonium-200 (Ch-200),


Mice

(Biswas and Khuda-Bukhsh 2002) BMC.ComplementAltern.Med 2:4.

Feeding of Chelidonium reduced genotoxic effects to a significant extent (p < 0.05 to p < 0.001).

Two potencies of Chelidonium (Ch-30, Ch-200)

p-dimethylaminoazobenzene (p-DAB)-induced hepatocarcinogenesis

Mice

(Chakrabarti, Biswas, and Khuda-Bukhsh2001) Indian J.Exp.Biol. 39 (12):1235-1242.

AMD had genotoxic effects of its own. Sonicated mice fed with Arnica 30 showed appreciably reduced genotoxicity

Actinomycin D or Arnica 30/placebo

Cytogenetical damage induced by exposures to ultrasonication

Mice

RefResultsInterventionDisease ModelAnimal

*The formula of Canova is composed of 19x Thuya occidentalis, 18x Bryonia alba, 11x Aconitum napellus, 19x Arsenicum album and 18x Lachesis muta(Viperidae) venom, all extracted and diluted in 70% alcohol, in equal parts.

(Sato et al. 2005) Homeopathy 94 (1):26-32.

Delay in the development, increased infiltration by lymphoid cells with active treatment. Increased number of leukocytes and lymphocytes.

Canova, a homeopathic complex medicine*

Sarcoma 180Mice

(Biswas et al. 2005) J Altern.Complement Med 11 (5):839-854.

Carcinosin 200 and Chelidonium 200 when administered alone show considerable ameliorative effect on cytogenetical endpoints and toxicity biomarkers

Carcinosin 200, fed alone and in combination with Chelidonium 200


Mice

(Datta, Biswas, and Khuda-Bukhsh 2004) Evid.Based.Complement Alternat.Med 1 (3):291-300.

Less chromosome aberrations in the drug-fed series. The amelioration by Merc Sol-200 appeared to be slightly more pronounced.

MercuriusSolubilis(Merc Sol-30 and Merc Sol-200)

Genotoxic Effects Produced by Mercuric Chloride

Mice

(Biswas and Khuda-Bukhsh 2004) Indian J.Exp.Biol. 42 (7):698-714.

Both Ch-30 and Ch-200 also modulated favourably some toxicity marker enzymes

Chelidonium-30 (Ch-30) and Chelidonium-200 (Ch-200),


Mice

(Biswas and Khuda-Bukhsh 2002) BMC.ComplementAltern.Med 2:4.

Feeding of Chelidonium reduced genotoxic effects to a significant extent (p < 0.05 to p < 0.001).

Two potencies of Chelidonium (Ch-30, Ch-200)

p-dimethylaminoazobenzene (p-DAB)-induced hepatocarcinogenesis

Mice

(Chakrabarti, Biswas, and Khuda-Bukhsh2001) Indian J.Exp.Biol. 39 (12):1235-1242.

AMD had genotoxic effects of its own. Sonicated mice fed with Arnica 30 showed appreciably reduced genotoxicity

Actinomycin D or Arnica 30/placebo

Cytogenetical damage induced by exposures to ultrasonication

Mice


4

Effects of homeopathic treatment on animal models of cancerogenesis (part 2)

(Pathak et al. 2006 and 2007) Mol.Cell Biochem. 285 (1-2):121-131.Forsch.Komplementarmed. 14 (3):148-156.

Protection from chromosome aberrations and toxicity biomarkers

Lycopodium 30c and 200C


Mice

(Bhattacharjee, Pathak, and Khuda-Bukhsh2007)eCAM Advance access

Less number of liver tumors and of chromosome aberrations reeducedtoxicity parameters

Natrum Sulphuricum200 (Nat Sulph-200)

HepatocarcinogenesisthroughP-DAB andPhenobarbital

Mice

(MacLaughlin et al. 2006)Integr.Cancer Ther. 5 (4):362-372.

Xenograft size was significantly reduced in Sabal serrulata-treated mice compared to untreated controls (P=.012). No effect was observed on breast tumor growth.

Sabal serrulata, Thuja occidentalis, and Conium maculatum

Human prostate xenografts cancer growth

Mice

(Kumar et al. 2007) Asian Pac.J Cancer Prev. 8 (1):98-102.

Homeopathic drugs retarded the tumor growth and reduced the marker enzymes (Ruta 200c of liver tumor, Ruta 200c and phosphorus 1M of sarcomas)

Ruta, Hydrastis, Lycopodium and Thuja (200C), Phosphorus 1M

N'-nitrosodiethylamine(NDEA) induced hepatocellularcarcinoma and sarcomas

Rats

(Banerjee et al. 2007) J Vet.Med.A PhysiolPathol.Clin.Med. 54 (7):370-376.

Drug fed mice showed reduced toxicity at statistically significant levels in respect of all the parameters studied

Arsenicum album 200alcohol (Alcohol-200)

Genotoxicity induced by repeated injections of arsenic trioxide

Mice

(Jonas et al. 2006)Integr.Cancer Ther. 5 (4):343-349.

23% reduction in tumor incidence (P < .0001), and 38% reduction in tumorvolume (P < .02). Tumors showed a 19% increase in apoptotic cell death (P < .05).

Thuja occ. 1000c, Conium mac. 1000c, Sabal serr. 200c, Carcinosin 1000c from MAT-LyLu cells

Prostate cancer (rats injected with MAT-LyLu cells)

Rats


(Pathak et al. 2006 and 2007) Mol.Cell Biochem. 285 (1-2):121-131.Forsch.Komplementarmed. 14 (3):148-156.

Protection from chromosome aberrations and toxicity biomarkers

Lycopodium 30c and 200C


Mice

(Bhattacharjee, Pathak, and Khuda-Bukhsh2007)eCAM Advance access

Less number of liver tumors and of chromosome aberrations reeducedtoxicity parameters

Natrum Sulphuricum200 (Nat Sulph-200)

HepatocarcinogenesisthroughP-DAB andPhenobarbital

Mice

(MacLaughlin et al. 2006)Integr.Cancer Ther. 5 (4):362-372.

Xenograft size was significantly reduced in Sabal serrulata-treated mice compared to untreated controls (P=.012). No effect was observed on breast tumor growth.

Sabal serrulata, Thuja occidentalis, and Conium maculatum

Human prostate xenografts cancer growth

Mice

(Kumar et al. 2007) Asian Pac.J Cancer Prev. 8 (1):98-102.

Homeopathic drugs retarded the tumor growth and reduced the marker enzymes (Ruta 200c of liver tumor, Ruta 200c and phosphorus 1M of sarcomas)

Ruta, Hydrastis, Lycopodium and Thuja (200C), Phosphorus 1M

N'-nitrosodiethylamine(NDEA) induced hepatocellularcarcinoma and sarcomas

Rats

(Banerjee et al. 2007) J Vet.Med.A PhysiolPathol.Clin.Med. 54 (7):370-376.

Drug fed mice showed reduced toxicity at statistically significant levels in respect of all the parameters studied

Arsenicum album 200alcohol (Alcohol-200)

Genotoxicity induced by repeated injections of arsenic trioxide

Mice

(Jonas et al. 2006)Integr.Cancer Ther. 5 (4):343-349.

23% reduction in tumor incidence (P < .0001), and 38% reduction in tumorvolume (P < .02). Tumors showed a 19% increase in apoptotic cell death (P < .05).

Thuja occ. 1000c, Conium mac. 1000c, Sabal serr. 200c, Carcinosin 1000c from MAT-LyLu cells

Prostate cancer (rats injected with MAT-LyLu cells)

Rats


5

Homeopathic medicine and animal (murine)cancer models: SUMMARY

{KUMAR2007} {PREETHI2006} Cancer Research Centre, Kerala State,

India

Inhibits liver tumor development(Ruta max effect in one paper and reported

as the only remedy in another)

Ruta, Hydrastis, Lycopodium , Thuja200CH, Phosphorus 1000CH

{MACLAUGHLIN2006}Georgetown University, Washington, DC

Reduces prostate cancer incidence and tumor volume

Sabal S. 200CH

{JONAS2006A}Samueli Institute, Alexandria, VA, USA


Thuja 1000CH, Conium 1000CH, Sabal S. 200CHCarcinosin 1000CH

{SATO2005}Universidade do Vale do Itajai, Brazil

Inhibit development of sarcomas-180Canova complex (Thuja , Bryonia, Aconitum,Arsenicum, Lachesis)

{BHATTACHARJEE2007} University of Kalyani, Nadia, India

Reduces p-DAB-induced tumorsincidence, genotoxicity and improves Biomarkers

Natrum sulf. 200CH

{PATHAK2006} {PATHAK2007} University of Kalyani, Nadia, India

Reduces p-DAB-induced tumorsincidence, genotoxicity and improvesBiomarkers

Lycopodium 30CH and 200CH

{DATTA2004} University of Kalyani, Nadia, India

Reduces mercury-induced genotoxicityMercurius Sol 30CH and200CH

{BISWAS2002} {BISWAS2004}{BISWAS2005}University of Kalyani, Nadia, India


Chelidonium 30CH and 200CH, alone or withCarcinosin 200CH

{CHAKRABARTI2001} University of Kalyani, Nadia, India

Reduces sonication-induced Genotoxicity

Arnica 30CH

Group and referenceEffectDrug

{KUMAR2007} {PREETHI2006} Cancer Research Centre, Kerala State,

India

Inhibits liver tumor development(Ruta max effect in one paper and reported

as the only remedy in another)

Ruta, Hydrastis, Lycopodium , Thuja200CH, Phosphorus 1000CH

{MACLAUGHLIN2006}Georgetown University, Washington, DC


Sabal S. 200CH

{JONAS2006A}Samueli Institute, Alexandria, VA, USA


Thuja 1000CH, Conium 1000CH, Sabal S. 200CHCarcinosin 1000CH

{SATO2005}Universidade do Vale do Itajai, Brazil

Inhibit development of sarcomas-180Canova complex (Thuja , Bryonia, Aconitum,Arsenicum, Lachesis)

{BHATTACHARJEE2007} University of Kalyani, Nadia, India


Natrum sulf. 200CH

{PATHAK2006} {PATHAK2007} University of Kalyani, Nadia, India

Reduces p-DAB-induced tumorsincidence, genotoxicity and improvesBiomarkers

Lycopodium 30CH and 200CH

{DATTA2004} University of Kalyani, Nadia, India

Reduces mercury-induced genotoxicityMercurius Sol 30CH and200CH

{BISWAS2002} {BISWAS2004}{BISWAS2005}University of Kalyani, Nadia, India


Chelidonium 30CH and 200CH, alone or withCarcinosin 200CH

{CHAKRABARTI2001} University of Kalyani, Nadia, India

Reduces sonication-induced Genotoxicity

Arnica 30CH

Group and referenceEffectDrug

PPT PPT –– 3.63.6

6

Effects of homeopathic preparations on human prostate cancer growth in cellular and animal modelsB. W. MacLaughlin et al. Integr. Cancer Ther., 5 (2006) 362-372.

PPT PPT –– 3.53.5

7

Effects of homeopathic treatment on cell models of cancerogenesis

(Preethi, Kuttan, and Kuttan 2006) Asian Pacific J Cancer Prev7:439-443.

Cytocidal action of both preparations

Ruta graveolens (MT and 200C)

Cell growthLymphomaAscites and

others

(MacLaughlin et al. 2006) Integr.CancerTher. 5 (4):362-372.

33% decrease ofprostate cell proliferation, noeffect on breast cells.

Sabal serrulata (sawpalmetto) 100CH

Cell proliferationHuman prostate cancer andbreast cancer cell lines.

(Thangapazham et al. 2006b) Integr.CancerTher. 5 (4):350-355.

No significant changes of anytreatment

Conium maculatum, Sabalserrulata, Thuja

occidentalis, and a MATLyLu Carcinosin nosode

mRNA levels of theapoptotic genes or thecytokines in prostatetumor

Copenhagen rattumor tissues

(Thangapazham et al.2006a) Integr.CancerTher. 5 (4):356-361.

No effects on cellviability or geneexpression

Conium maculatum, Sabalserrulata, Thujaoccidentalis, Asterias,Phytolacca, and Carcinosinin several potencies

Tumor cell viability andapoptosis geneexpression

Prostate andbreast cancercell lines

(Jonas et al. 2006)Integr.Cancer Ther. 5(4):343-349.


Thuja occidentalis 1000c,Conium maculatum 1000c,Sabal serrulata 200c


Prostate tumorcells

(Nadareishvili 2006)Georgian.Med.News(136):99-101.

Stimulates ionictransport andNa,K-ATPase

Phosphoric acid (PA), atlow (10(-14) and high

10(-42)) dilutions

Continuous recording of Na(+), K(+), and Ca(2+)with selective electrodes

Ehrlichcarcinoma

(Walchli, Baumgartner,and Bastide 2006). JAltern. ComplementMed. 12 (5):421-427.

Increased cellviability in primarylymphocytes.Weaker effect incancerous cells

Pretreatment with either low concentrations (nM-microM)or high potencies (15-20c)of cadmium (Isopatic)

Cell viabilityNormal and leukemiclymphocytes

RefResultsInterventionLaboratory testCells

(Preethi, Kuttan, and Kuttan 2006) Asian Pacific J Cancer Prev7:439-443.

Cytocidal action of both preparations

Ruta graveolens (MT and 200C)

Cell growthLymphomaAscites and

others

(MacLaughlin et al. 2006) Integr.CancerTher. 5 (4):362-372.

33% decrease ofprostate cell proliferation, noeffect on breast cells.

Sabal serrulata (sawpalmetto) 100CH

Cell proliferationHuman prostate cancer andbreast cancer cell lines.

(Thangapazham et al. 2006b) Integr.CancerTher. 5 (4):350-355.

No significant changes of anytreatment

Conium maculatum, Sabalserrulata, Thuja

occidentalis, and a MATLyLu Carcinosin nosode

mRNA levels of theapoptotic genes or thecytokines in prostatetumor

Copenhagen rattumor tissues

(Thangapazham et al.2006a) Integr.CancerTher. 5 (4):356-361.


Conium maculatum, Sabalserrulata, Thujaoccidentalis, Asterias,Phytolacca, and Carcinosinin several potencies


Prostate andbreast cancercell lines

(Jonas et al. 2006)Integr.Cancer Ther. 5(4):343-349.


Thuja occidentalis 1000c,Conium maculatum 1000c,Sabal serrulata 200c


Prostate tumorcells

(Nadareishvili 2006)Georgian.Med.News(136):99-101.

Stimulates ionictransport andNa,K-ATPase

Phosphoric acid (PA), atlow (10(-14) and high

10(-42)) dilutions

Continuous recording of Na(+), K(+), and Ca(2+)with selective electrodes

Ehrlichcarcinoma

(Walchli, Baumgartner,and Bastide 2006). JAltern. ComplementMed. 12 (5):421-427.

Increased cellviability in primarylymphocytes.Weaker effect incancerous cells

Pretreatment with either low concentrations (nM-microM)or high potencies (15-20c)of cadmium (Isopatic)

Cell viabilityNormal and leukemiclymphocytes

RefResultsInterventionLaboratory testCells

8

EXAMPLES OF HIGH-DILUTION EXPERIMENTS

Chirumbolo 1993Inhibition of superoxide12 x to 30 xMagnesium phosPhosphorus

Human neutrophils

Scherr 2006Affect growth kinetics9-30 CHAxoxystrobinPhosphorus

Saccharomyces

Marotta 2002Increases viability10-27CycloheximideNeurons

Belon 1999-2006 et al (including Verona Group)

Inhibition10-24

��10-32

HistamineHuman basophils

Oberbaum 1998Speeds up wound healing10-60SilicaMouse ears

Cristea 1991-98Increases Ach-induced spasmIncreases Ach-induced spasm

60 CH200 CH

Atropa belladonnaRat duodenum

Sukul 1991-98Reduces firing rate in rats under high-salt diet

10-60Sodium chlorideRat Hypothalamus

ArsenicSilver nitrate

Acetylsalicylic acid

Nux vomica

Bursin

IgE

Agent

Betti 1997/2001Pongratz 1998

Protect from toxicityEnhances growth

10-45

26 D

Wheat germination

Doutremepuich 1998Pro-thrombotic10-30Mouse blood

Sukul 1999Reduction of alchol-induced sleep time30 CHMice nervous system

Youbicier-Simo 1993-97Immunomodulatory and endocrine activity15 CH10-27g

Cicken embrio

Davenas 1988Stimulation (not confirmed by others)10-60Human basophils

Ref.EffectDilutionSystem

Chirumbolo 1993Inhibition of superoxide12 x to 30 xMagnesium phosPhosphorus

Human neutrophils

Scherr 2006Affect growth kinetics9-30 CHAxoxystrobinPhosphorus

Saccharomyces

Marotta 2002Increases viability10-27CycloheximideNeurons

Belon 1999-2006 et al (including Verona Group)

Inhibition10-24

��10-32

HistamineHuman basophils

Oberbaum 1998Speeds up wound healing10-60SilicaMouse ears

Cristea 1991-98Increases Ach-induced spasmIncreases Ach-induced spasm

60 CH200 CH

Atropa belladonnaRat duodenum

Sukul 1991-98Reduces firing rate in rats under high-salt diet

10-60Sodium chlorideRat Hypothalamus

ArsenicSilver nitrate

Acetylsalicylic acid

Nux vomica

Bursin

IgE

Agent

Betti 1997/2001Pongratz 1998

Protect from toxicityEnhances growth

10-45

26 D

Wheat germination

Doutremepuich 1998Pro-thrombotic10-30Mouse blood

Sukul 1999Reduction of alchol-induced sleep time30 CHMice nervous system

Youbicier-Simo 1993-97Immunomodulatory and endocrine activity15 CH10-27g

Cicken embrio

Davenas 1988Stimulation (not confirmed by others)10-60Human basophils

Ref.EffectDilutionSystem

9

FARMACOLOGIA DELLE ALTE DILUIZIONIStudi sulle basi biochimiche e fisiche del medicinale omeopatico

Sukul N.C., Sukul A.

EDITORE: Salus Infirmorum Pad ova

[email protected] 049-755446

1. Modi per preparare i rimedi omeopatici e per conserva rli.2. Evidenze cliniche ottenute a sostegno delle alte diluizion i

nell’uomo, unitamente ad esperimenti di laboratorio3. Caratteristiche fisiche dei medicamenti in diluizioni u ltra-

alte (risonanza magnetica nucleare e spettriall’infrarosso).

4. Possibili meccanismi d’azione delle alte diluizioni suisistemi viventi.

10

Basic research in homeopathy

TYPICAL CELLULAR MODELS FROM THE LITERATURE

• Lymphocytes (Colas, Bildet, Bastide, Wagner, Chirila)

• Basophils (Poitevin, Sainte Laudy, Belon, Davenas, Ovelgonne, Hirst, Mannaioni, Ennis)

• Granulocytes (Poitevin, Fletcher, Chirumbolo, Bellavite)

• Cell lines: Fibroblasts (Boiron, Mansvelt, Fougeray) Renal cells(Delbancut), Hepatoma (van Wijk, Wiegant), Neurons (Jonas)

• Vegetable cells (Guillemain, Bornoroni, Betti)

• Lymphocytes (Colas, Bildet, Bastide, Wagner, Chirila)

• Basophils (Poitevin, Sainte Laudy, Belon, Davenas, Ovelgonne, Hirst, Mannaioni, Ennis)

• Granulocytes (Poitevin, Fletcher, Chirumbolo, Bellavite)

• Cell lines: Fibroblasts (Boiron, Mansvelt, Fougeray) Renal cells(Delbancut), Hepatoma (van Wijk, Wiegant), Neurons (Jonas)

• Vegetable cells (Guillemain, Bornoroni, Betti)

11

RestingResting cellcell

AntiAnti--IgEIgE(Medium (Medium dosesdoses))AntiAnti--IgEIgE(Medium (Medium dosesdoses))

* *** **

UltraUltra--high dilution high dilution (10(10--2222, 10, 10--3434 M)M)of of HistamineHistamine

* UltraUltra--high dilution high dilution (10(10--2222, 10, 10--3434 M)M)of of HistamineHistamine

*

InhibitionInhibition of of responseresponsetoto AntiAnti--IgEIgE

** *

*


** *

*


** *

*

CONFIRMATION OF PHENOMENON WITH CD63 EXPRESSION(S.LAUDY, BELON ET AL., 1996-2006, BROWN AND ENNIS, 2001)

PPT PPT –– 3.53.5

12

HISTAMINE DILUTIONS MODULATE BASOPHIL ACTIVATION

� Study 1 used a blinded multi-centre approach in 4 centres. Study 2, related to the confirmation of the multi-centre study by flow cytometry, was performed independently in 3 laboratories. Study 3 examined the histamine release (one laboratory) and the activity of H(2) receptor antagonists and structural analogues (two laboratories).

� RESULTS: High dilutions of histamine (10-30-10-38 M) influence the activation of human basophils measured by alcian blue staining. The degree of inhibition depends on the initial level of anti-IgE induced stimulation, with the greatest inhibitory effects seen at lower levels of stimulation. This multicentre study was confirmed in the three laboratories by using flow cytometry and in one laboratory by histamine release.

� Inhibition of basophil activation by high dilutions of histamine was reversed by anti-H2 and was not observed with histidine these results being in favour of the specificity of this effect

� We are however unable to explain our findings and are reporting them toencourage others to investigate this phenomenon.

� Study 1 used a blinded multi-centre approach in 4 centres. Study 2, related to the confirmation of the multi-centre study by flow cytometry, was performed independently in 3 laboratories. Study 3 examined the histamine release (one laboratory) and the activity of H(2) receptor antagonists and structural analogues (two laboratories).

� RESULTS: High dilutions of histamine (10-30-10-38 M) influence the activation of human basophils measured by alcian blue staining. The degree of inhibition depends on the initial level of anti-IgE induced stimulation, with the greatest inhibitory effects seen at lower levels of stimulation. This multicentre study was confirmed in the three laboratories by using flow cytometry and in one laboratory by histamine release.

� Inhibition of basophil activation by high dilutions of histamine was reversed by anti-H2 and was not observed with histidine these results being in favour of the specificity of this effect

� We are however unable to explain our findings and are reporting them toencourage others to investigate this phenomenon.

Belon P, Cumps J, Ennis M, Mannaioni PF, Roberfroid M, Sainte-Laudy J, Wiegant FA. - Inflamm Res. 2004 May;53(5):181-8.

PPT – 3.5, 3.9

13

Improvement of flow cytometric analysis of basophil activation inhibition by high histamine dilutions. A novel basophil specific marker: CD 203c.

• Sainte-Laudy J, Belon P. • Homeopathy. 2006 Jan;95(1):3-8.

We investigated if the use of CD 203c, a basophil specific, earlier marker than CD 63 of the activation cascade, increased the sensitivity of the method, testing two target histamine dilutions, 10(-4) (2C) and 10(-32) M (16C).

• Basophils, obtained from buffy coats, were pre-incubated with the histamine dilutions and activated by two agonists: anti-IgE and fMLP (formyl-methionyl-leucyl-phenylalanine peptide).

• The cells were labelled with anti-IgE, anti-CD 13 and anti-CD 14 for basophil selection, and anti-CD 63 and anti-CD 203c for basophil activation. Results were expressed in up-regulation percentage for CD 63 or mean intensity of fluorescence (MFI) for CD 203c.

• RESULTS: Histamine 10(-4) M (2C) and histamine 10(-32) M (16C) were capable of inhibiting both IgE-dependent (anti-IgE) and IgE-independent (fMLP) basophil activation. The percentage inhibition depended on the activation marker used. The highest inhibition for histamine dilution 16C was observed with CD 203c (38%, P<0.001), approximately half the inhibition observed with histamine 2C (73%).

• CONCLUSION: The use of CD 203c instead of CD 63 increased the magnitude of the response.

14

Dilutions (CH)2 10 11 12 13 14 15 16

MFI (

% o

f ant

i-IgE

con

trol

)

0

50

100

150

200

CD203c-PE

071016-riassuntivo Cd203c vs water.jnb

Dilutions (CH)2 10 11 12 13 14 15 16

MFI (

% o

f ant

i-IgE

con

trol

)

0

50

100

150

200

CD203c-PEPURE WATER(diluted and succussed)

HISTAMINE DIHYDROCHLORIDE(diluted and succussed)

Untreatedcontrol

Untreatedcontrol

EFFECTS OF HISTAMINE DILUTIONS ON HUMAN BASOPHIL ACTIVATION

© P. Bellavite, Università di Verona

This study is supportedby a research grant

from Boiron s.r.l. toVerona University

EFFECTS OF HISTAMINE DILUTIONS/DYNAMIZATIONS (POTENCIES) ON THE HUMAN BASOPHIL ACTIVATION IN VITRO (CD203 expression)

RECENT (UNPUBLISHED) EVIDENCE FROM VERONA’S LAB (Bellavite, Chirumbolo, Ortolani, Vella)

Nome file: BELLAVITE-relazione.doc Directory: C:\Documents and

Settings\Mauri\Documenti\allproject\web\omeopatia.org\_URGENTE Modello: C:\Documents and Settings\Mauri\Dati

applicazioni\Microsoft\Modelli\Normal.dot Titolo: Oggetto: Autore: Utente Windows Parole chiave: Commenti: Data creazione: 15/11/2007 10.48 Numero revisione: 2 Data ultimo salvataggio: 15/11/2007 10.48 Autore ultimo salvataggio: SCUOLA MEDICINA OMEOPATICA Tempo totale modifica0 minuti Data ultima stampa: 04/12/2007 18.05 Come da ultima stampa completa Numero pagine: 14 Numero parole: 6 (circa) Numero caratteri: 35 (circa)